The Effect of Calcium Chloride on Alleviation of in vitro Shoot Tip Necrosis of Elsholtzia stachyodes
Abstract
Elsholtzia stachyodes is classified in the family of Lamiaceae. Its leaves are used as vegetables by hill tribe people in Thailand. This plant is interesting since it contains high content of flavonoids which possess anticancer, antioxidant, and anti-inflammatory activities. However, seed propagation success is limited by low viability and germination of seed. Therefore, tissue culture technique is an alternative method to reach high quantity of the plant within a short period of time. For shoot multiplication, shoot tip necrosis is a major problem for E. stachyodes. Therefore, the current research aims to study effects of calcium chloride on reducing shoot tip necrosis by culturing the 4 week-old shoots of E. stachyodes on MS medium containing 440 mg/L of calcium chloride and the modified MS media containing 880 or 1320 mg/L of calcium chloride in combination with BA at 0, 0.5, or 1 mg/L for 4 weeks. The results showed that the most appropriate medium formula was the modified MS media containing 880 mg/mL of calcium chloride in combination with 0.5 mg/L of BA. This formulation resulted in 100% shoot induction with 76.34 % of normal shoot characteristics, and the average number of shoots was 2.36 shoots/explant. Keywords : tissue culture, Elsholtzia stachyodes, shoot tip necrosis, calcium chloride, BAReferences
Bairu, M. W., Jain, N., Stirk, W. A., Doležal, K., & Van Staden, J. (2009). Solving the problem of shoot-tip necrosis in
Harpagophytum procumbens by changing the cytokinin types, calcium and boron concentrations in the
medium. South African Journal of Botany, 75(1), 122-127.
Barghchi, M., & Alderson, P. G. (1996). The control of shoot tip necrosis in Pistacia vera L. in vitro. Plant Growth
Regulation, 20(1), 31-35.
Bongcheewin, B., & Chantaranothai, P. (2008). Two New Records of Elsholtzia Willd.(Lamiaceae) for
Thailand. Tropical Natural History, 8(1), 1-5.
Bongcheewin, B., Chantaranothai, P., & Paton, A. (2015). Elsholtzia (Lamiaceae) in Thailand. Blumea-Biodiversity,
Evolution and Biogeography of Plants, 59(3), 209-214.
Chiruvella, K. K., Mohammed, A., Dampuri, G., & Ghanta, R. G. (2011). In vitro shoot regeneration and control of
shoot tip necrosis in tissue cultures of Soymida febrifuga (Roxb.) A. Juss. Plant Tissue Culture and Biotechnology, 21(1), 11-25.
De Block, M. (1990). Factors influencing the tissue culture and the Agrobacterium tumefaciens-mediated
transformation of hybrid aspen and poplar clones. Plant Physiology, 93(3), 1110-1116.
George, E. F., Hall, M. A., & De Klerk, G. J. (2008). Plant growth regulators II: cytokinins, their analogues and
Antagonists, Plant propagation by tissue culture. (pp. 205-226). Dordrecht: Springer.
Harmon, A. C., Gribskov, M., Gubrium, E., & Harper, J. F. (2001). The CDPK superfamily of protein kinases.
New Phytologist, 151(1), 175-183.
Hepler, P. K. (2005). Calcium: a central regulator of plant growth and development. The Plant Cell, 17(8),
2142-2155.
Jain, N., Bairu, M. W., Stirk, W. A., & Van Staden, J. (2009). The effect of medium, carbon source and explant on
regeneration and control of shoot-tip necrosis in Harpagophytum procumbens. South African Journal of
Botany, 75(1), 117-121.
Khomdram, S. D., Salam, J. S., & Singh, P. K. (2011). Estimation of nutritive indices in eight Lamiaceae plants of
Manipur. Am. J. Food Technol, 6(10), 924-931.
Khomdram, S. D., & Singh, P. K. (2011). Polyphenolic compounds and free radical scavenging activity in eight
Lamiaceae herbs of Manipur. Notulae Scientia Biologicae, 3(2), 108-113.
Kishore, K., Patnaik, S., & Shukla, A. K. (2015). Optimization of method to alleviate in vitro shoot tip necrosis in
Trichosanthes dioica Roxb. Indian Journal of Biotechnology, 14, 107-111.
Kumari, M., Agnihotri, D., Chanotiya, C. S., Mathur, A. K., Lal, R. K., & Mathur, A. (2017). Chemical and genetic
stability of methyl chavicol-rich Indian basil (Ocimum basilicum var. CIM-Saumya) micropropagated
in vitro. South African Journal of Botany, 113, 186-191.
Machado, M. P., Silva, A. L. L. D., Biasi, L. A., Deschamps, C., Bespalhok Filho, J. C., & Zanette, F. (2014).
Influence of calcium content of tissue on hyperhydricity and shoot-tip necrosis of in vitro regenerated
shoots of Lavandula angustifolia Mill. Brazilian Archives of Biology and Technology, 57(5), 636-643.
Martin, K. P., Zhang, C. L., Slater, A., & Madassery, J. (2007). Control of shoot necrosis and plant death during
micro-propagation of banana and plantains (Musa spp.). Plant Cell, Tissue and Organ Culture, 88(1),
51-59.
Miyazawa, M., Okuno, Y., Nakamura, S. I., & Kosaka, H. (2000). Antimutagenic activity of flavonoids from
Pogostemon cablin. Journal of agricultural and food chemistry, 48(3), 642-647.
Murashige, T., & Skoog, F. (1962). A revised medium for rapid growth and bio assays with tobacco tissue
cultures. Physiologia plantarum, 15(3), 473-497.
Piagnani, C., Zocchi, G., & Mignani, I. (1996). Influence of Ca2+ and 6-benzyladenine on chestnut (Castanea
sativa Mill.) in vitro shoot-tip necrosis. Plant Science, 118(1), 89-95.
Prakash, S., & Van Staden, J. (2007). Micropropagation of Hoslundia opposita Vahl—a valuable medicinal
plant. South African Journal of Botany, 73(1), 60-63.
Raven, J. A. (1977). H+ and Ca2+ in phloem and symplast: relation of relative immobility of the ions to the
cytoplasmic nature of the transport paths. New Phytologist, 79(3), 465-480.
Saha, S., Dey, S. T., Adhikari, S., & Ghosh, P. D. (2010). In vitro multiple shoot regeneration and analysis of
genetic fidelity of Mentha piperita L. Bionature, 30(2), 71-81.
Saha, S., Kader, A., Sengupta, C., & Ghosh, P. (2012). In vitro propagation of Ocimum gratissimum L.
(Lamiaceae) and its evaluation of genetic fidelity using RAPD marker. Am J Plant Sci, 3(1), 64-74.
Saha, S., Roy, S., Sengupta, C., & Ghosh, P. (2014). Micropropagation and analysis of genetic stability in
regenerated plantlets of Ocimum canum Sims. Indian Journal of Plant Physiology, 19(2), 174-183.
Sita, G. L., & Swamy, B. R. (1993). Regeneration of plantlets from leaf disc cultures of rosewood: control of leaf
abscission and shoot tip necrosis. Plant Science, 88(1), 107-112.
Trisonthi, C., & Trisonthi, P. (2009). Ethnobotanical study in Thailand, a case study in Khun Yuam District
Maehongson Province. Thai Journal of Botany, 1(1), 1-23. (in Thai)
Van Doorn, W. G., Beers, E. P., Dangl, J. L., Franklin-Tong, V. E., Gallois, P., Hara-Nishimura, I., & Mur, L. A. J.
(2011). Morphological classification of plant cell deaths. Cell death and differentiation, 18(8), 1241-1246.
Zhang, D. H., Wadsworth, P., & Hepler, P. K. (1992). Modulation of anaphase spindle microtubule structure in
stamen hair cells of Tradescantia by calcium and related agents. Journal of Cell Science, 102(1), 79-89.
Zhang, L., & Lu, Y. T. (2003). Calmodulin-binding protein kinases in plants. Trends in plant science, 8(3), 123-127.
Harpagophytum procumbens by changing the cytokinin types, calcium and boron concentrations in the
medium. South African Journal of Botany, 75(1), 122-127.
Barghchi, M., & Alderson, P. G. (1996). The control of shoot tip necrosis in Pistacia vera L. in vitro. Plant Growth
Regulation, 20(1), 31-35.
Bongcheewin, B., & Chantaranothai, P. (2008). Two New Records of Elsholtzia Willd.(Lamiaceae) for
Thailand. Tropical Natural History, 8(1), 1-5.
Bongcheewin, B., Chantaranothai, P., & Paton, A. (2015). Elsholtzia (Lamiaceae) in Thailand. Blumea-Biodiversity,
Evolution and Biogeography of Plants, 59(3), 209-214.
Chiruvella, K. K., Mohammed, A., Dampuri, G., & Ghanta, R. G. (2011). In vitro shoot regeneration and control of
shoot tip necrosis in tissue cultures of Soymida febrifuga (Roxb.) A. Juss. Plant Tissue Culture and Biotechnology, 21(1), 11-25.
De Block, M. (1990). Factors influencing the tissue culture and the Agrobacterium tumefaciens-mediated
transformation of hybrid aspen and poplar clones. Plant Physiology, 93(3), 1110-1116.
George, E. F., Hall, M. A., & De Klerk, G. J. (2008). Plant growth regulators II: cytokinins, their analogues and
Antagonists, Plant propagation by tissue culture. (pp. 205-226). Dordrecht: Springer.
Harmon, A. C., Gribskov, M., Gubrium, E., & Harper, J. F. (2001). The CDPK superfamily of protein kinases.
New Phytologist, 151(1), 175-183.
Hepler, P. K. (2005). Calcium: a central regulator of plant growth and development. The Plant Cell, 17(8),
2142-2155.
Jain, N., Bairu, M. W., Stirk, W. A., & Van Staden, J. (2009). The effect of medium, carbon source and explant on
regeneration and control of shoot-tip necrosis in Harpagophytum procumbens. South African Journal of
Botany, 75(1), 117-121.
Khomdram, S. D., Salam, J. S., & Singh, P. K. (2011). Estimation of nutritive indices in eight Lamiaceae plants of
Manipur. Am. J. Food Technol, 6(10), 924-931.
Khomdram, S. D., & Singh, P. K. (2011). Polyphenolic compounds and free radical scavenging activity in eight
Lamiaceae herbs of Manipur. Notulae Scientia Biologicae, 3(2), 108-113.
Kishore, K., Patnaik, S., & Shukla, A. K. (2015). Optimization of method to alleviate in vitro shoot tip necrosis in
Trichosanthes dioica Roxb. Indian Journal of Biotechnology, 14, 107-111.
Kumari, M., Agnihotri, D., Chanotiya, C. S., Mathur, A. K., Lal, R. K., & Mathur, A. (2017). Chemical and genetic
stability of methyl chavicol-rich Indian basil (Ocimum basilicum var. CIM-Saumya) micropropagated
in vitro. South African Journal of Botany, 113, 186-191.
Machado, M. P., Silva, A. L. L. D., Biasi, L. A., Deschamps, C., Bespalhok Filho, J. C., & Zanette, F. (2014).
Influence of calcium content of tissue on hyperhydricity and shoot-tip necrosis of in vitro regenerated
shoots of Lavandula angustifolia Mill. Brazilian Archives of Biology and Technology, 57(5), 636-643.
Martin, K. P., Zhang, C. L., Slater, A., & Madassery, J. (2007). Control of shoot necrosis and plant death during
micro-propagation of banana and plantains (Musa spp.). Plant Cell, Tissue and Organ Culture, 88(1),
51-59.
Miyazawa, M., Okuno, Y., Nakamura, S. I., & Kosaka, H. (2000). Antimutagenic activity of flavonoids from
Pogostemon cablin. Journal of agricultural and food chemistry, 48(3), 642-647.
Murashige, T., & Skoog, F. (1962). A revised medium for rapid growth and bio assays with tobacco tissue
cultures. Physiologia plantarum, 15(3), 473-497.
Piagnani, C., Zocchi, G., & Mignani, I. (1996). Influence of Ca2+ and 6-benzyladenine on chestnut (Castanea
sativa Mill.) in vitro shoot-tip necrosis. Plant Science, 118(1), 89-95.
Prakash, S., & Van Staden, J. (2007). Micropropagation of Hoslundia opposita Vahl—a valuable medicinal
plant. South African Journal of Botany, 73(1), 60-63.
Raven, J. A. (1977). H+ and Ca2+ in phloem and symplast: relation of relative immobility of the ions to the
cytoplasmic nature of the transport paths. New Phytologist, 79(3), 465-480.
Saha, S., Dey, S. T., Adhikari, S., & Ghosh, P. D. (2010). In vitro multiple shoot regeneration and analysis of
genetic fidelity of Mentha piperita L. Bionature, 30(2), 71-81.
Saha, S., Kader, A., Sengupta, C., & Ghosh, P. (2012). In vitro propagation of Ocimum gratissimum L.
(Lamiaceae) and its evaluation of genetic fidelity using RAPD marker. Am J Plant Sci, 3(1), 64-74.
Saha, S., Roy, S., Sengupta, C., & Ghosh, P. (2014). Micropropagation and analysis of genetic stability in
regenerated plantlets of Ocimum canum Sims. Indian Journal of Plant Physiology, 19(2), 174-183.
Sita, G. L., & Swamy, B. R. (1993). Regeneration of plantlets from leaf disc cultures of rosewood: control of leaf
abscission and shoot tip necrosis. Plant Science, 88(1), 107-112.
Trisonthi, C., & Trisonthi, P. (2009). Ethnobotanical study in Thailand, a case study in Khun Yuam District
Maehongson Province. Thai Journal of Botany, 1(1), 1-23. (in Thai)
Van Doorn, W. G., Beers, E. P., Dangl, J. L., Franklin-Tong, V. E., Gallois, P., Hara-Nishimura, I., & Mur, L. A. J.
(2011). Morphological classification of plant cell deaths. Cell death and differentiation, 18(8), 1241-1246.
Zhang, D. H., Wadsworth, P., & Hepler, P. K. (1992). Modulation of anaphase spindle microtubule structure in
stamen hair cells of Tradescantia by calcium and related agents. Journal of Cell Science, 102(1), 79-89.
Zhang, L., & Lu, Y. T. (2003). Calmodulin-binding protein kinases in plants. Trends in plant science, 8(3), 123-127.
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2018-12-03
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